Adjustment of Codon Usage Frequencies by Codon Harmonization Improves Protein Expression and Folding
Adjustment of Codon Usage Frequencies by Codon Harmonization Improves Protein Expression and Folding
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DOI：
10.1007/978-1-61737-967-3_1
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发表时间：
2011-01-01
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影响因子：
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通讯作者：
Mease, Ryan M.
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文献类型：
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作者：
Angov, Evelina;Legler, Patricia M.;Mease, Ryan M.
在过去的二十年中，原核表达系统已被广泛用于许多治疗性蛋白质的生物生产。大部分的成功可以归因于原核蛋白质翻译和蛋白质折叠的基本原理的实施，以解决异源表达的问题（例如密码子使用替换、tRNA异源受体共表达、伴侣蛋白共表达）;然而，在异源宿主中的表达仍然是经验过程。为了进一步改善异源蛋白表达，我们开发了一种称为“密码子协调”的算法，其最接近来自天然宿主的密码子使用频率，并调整这些用于异源系统。这种方法的成功可能是由于翻译过程中蛋白质折叠的改善。虽然到目前为止只适用于大肠杆菌，密码子协调可以提供一个通用的策略，以提高可溶性的，功能性蛋白质的表达在异源宿主表达。
Over the past two decades, prokaryotic expression systems have been widely exploited for the bioproduction of many therapeutic proteins. Much of the success can be attributed to the implementation of basic principles of prokaryotic protein translation and protein folding to the problems of heterologous expression (e.g. codon usage substitutions, tRNA isoacceptor co-expression, chaperone co-expression); however, expression in a heterologous host still remains an empirical process. To improve heterologous protein expression further we have developed an algorithm termed "codon harmonization" that best approximates codon usage frequencies from the native host and adjusts these for use in the heterologous system. The success of this methodology may be due to improved protein folding during translation. Although so far exclusively applied to Escherichia coli, codon harmonization may provide a general strategy for improving the expression of soluble, functional proteins during heterologous host expression.